Analysis about ultrasonic misdiagnosis of lower extremity venous thrombosis

目的通过对下肢静脉血栓超声患者的超声资料进行回顾性分析,总结分析误诊漏诊的原因方法对70例临床怀疑为下肢静脉血栓的病人进行常规静脉彩超检查,对确诊为下肢静脉血栓的患者再行CT静脉造影检查,而后对两者诊断结果进行对照分析。结果 70例临床疑似为下肢静脉血栓病人中,确诊率为57.1%（40/70）,超声误诊率32%（8/25）。结论超声检查无创伤、重复性高,可准确诊断下肢静脉血栓形成。Objective The data of patients with lower extremity venous thrombosis were analyzed retrospectively, and the causes of misdiagnosis and missed diagnosis were summarized. Methods 70 patients with clinical suspicion of lower extremity venous thrombosis were treated by routine venous color Doppler ultrasonography, and the venous thrombosis was diagnosed by CT venous angiography.Results Among 700 patients with clinical suspicion of lower extremity venous thrombosis, the accuracy rate was 57.1%, and the misdiagnosis rate was 32%. Conclusion Because ultrasound examination has high repeatability and no trauma, it can be used to diagnose exactly the lower extremity venous thrombosis.昌吉回族自治州科技局项目（TG-201405

3种养殖方式下沐浴角骨海绵生长状况的差异分析

沐浴角骨海绵（Spongia officinalis）主要分布于海南沿海，其骨架不由骨针而是由海绵硬蛋白组成，因而拥有良好的吸水性和柔性，在海洋生物材料开发领域具有较好前景。为了培育该地域特色的海绵品种，在海南省儋州水牛坡村潮间带海域（109.40° E, 19.87° N）开展了为期一年半的沐浴角骨海绵人工养殖，通过组织块的长、宽、高变化表征海绵的增长，分析了3种养殖方式下该海绵的生长状况差异。结果显示：在瓷砖法、穿绳法和弹簧法养殖方式下的角骨海绵均有较高的存活率，分别为80%，60%和50%，其中以瓷砖法最佳。在瓷砖法下，海绵组织块的平均长度增长率为67.1%，显著高于穿绳法和弹簧法（p<0.05），平均宽度增长率为63.5%，平均高度增长率为73.2%，可见瓷砖法相对于其他两种方式在存活率和增长率有较大优势，适宜于沐浴角骨海绵的养殖。此外，生长指标与海水温度的相关性分析结果表明，沐浴角骨海绵组织块的增长与水温之间的相关性不显著，增长速度不呈季节性变化。综上，在自然条件下获得的沐浴角骨海绵的第一手养殖资料有助于基础的海绵生物学研究，促进未来当地特色海洋经济的发展。海南省本级部门预算项目厦门南方海洋研究中心项目(17GYY008NF04

Construction of eukaryotic vector pcDNA3.1-flag-pygo2 and expression in C6 cell

目的构建pcDNA3.1-flag-pygo2真核表达载体并在C6细胞中进行表达。方法从小鼠脑胶质细胞中提取总RNA,RT-PCR法反转录合成c DNA,设计引物,调取目的片段,与pcDNA3.1-flag载体连接后转化大肠杆菌DH5α,LB平板筛选菌落,提取质粒。重组质粒pcDNA 3.1-flag-pygo2经过酶切鉴定及测序后,阳离子脂质体法转染C6细胞并经免疫细胞化学染色及蛋白印迹检测重组体的表达。结果重构质粒pcDNA 3.1-flag-pygo2经限制性核酸内切酶EcoRⅠ,HindⅢ酶切分析及测序检查,表明真核表达载体构建正确;瞬时转染C6细胞后,免疫细胞荧光染色及蛋白印迹检测表明转染细胞能够表达外源Pygo2基因。结论成功构建了pcDNA3.1-flag-pygo2真核表达载体并能够在真核细胞中进行表达,这为今后研究pygo2基因在胶质瘤中的作用机制奠定了基础。Objective To construct the eukaryotic expression vector pcDNA3.1-flag-pygo2 and express the combined protein in C6 cell line.Methods To extract total RNA from primary glial cell of mouse and to synthesize c DNA by RT-PCR,then design primer and clone whole segment of pygo2 gene.After the targeted gene was inserted into vector pcDNA3.1-flag,the recombined plamid was transformed into E coli DH5α for LB agar plate screening and the recombined plasmid were extracted and purified.After verification by double enzyme digestion and sequencing.,the constructed eukaryotic expression plamid was transfected to C6 cell line by lipofectamin method,finally the protein expression was detected by immunocytochemical staining as well as western blot.Results The new constructed vector was confirmed by restricted enzyme Eco R I,HindIII digestion assay and correct Pygo2 was verified by sequenceing.finally,pcDNA3.1-flag-hpygo2 can express exogenous pygo2 gene in glioma C6 cell line after transient transfection by the determination of immunocytofluorescent staining and western blot.Conclusion The new plamid pcDNA3.1-flag-pygo2 was constructed successfully,and can express fused protein in eukaryotic cell,which establish the foundation for future research on pygo2 gene function in human glioma

Over-expression of Pygo2 Promotes C6 Cells Proliferation of Glioma

目的通过构建过表达PygO2的重组体上调PygO2表达,探讨其在大鼠胶质瘤C6细胞增殖中的作用及机制。方法重组体经ECOr I和HIndⅢ双酶切鉴定和dnA测序后,用脂质体2000将其转染大鼠胶质瘤C6细胞,采用WESTErn blOT检测外源PygO2蛋白表达,应用克隆形成实验和MTT法检测细胞增殖,流式细胞术检测细胞周期,采用WESTErn blOT检测过表达PygO2对C6细胞中CyClInd1、β-CATEnIn水平的影响,并采用细胞免疫荧光法检测其对C6细胞中CyClInd1、β-CATEnIn亚细胞定位的影响。结果双酶切和测序鉴定结果证实插入序列正确,重组体能有效上调PygO2表达。将重组体转染C6细胞上调PygO2表达后,细胞的生长增殖被显著促进,克隆形成显著增多,细胞周期进程从g1期至S期转变显著增强;且CyClInd1水平随之增高,亚定位无改变,β-CATEnIn水平和亚细胞定位无明显改变。结论成功构建了过表达PygO2的重组体,过表达PygO2通过增高CyClInd1水平,促进细胞从g1期进入S期,从而促进大鼠胶质瘤C6细胞增殖。Objective To up-regulate expression of Pygopus2(Pygo2) by construction of the recombinant vectors of over-expression of Pygo2 protein,and to explore the role and mechanism of over-expression of Pygo2 in C6 cells proliferation of glioma.Methods The recombinant plasmids were digested with EcoRⅠ and Hind Ⅲ to execute the restriction endonuclease identification,then the sequence analysis was assayed by DNA sequencing.The recombinant plasmids were transfected into cultured glioblastoma C6 cells using lipofectamineTM 2000.The exogenous Pygo2 protein level of C6 cells was detected by Western blot analysis.Colony forming assay and MTT assay were used to detect the cell proliferation,and cell cycle analysis was performed by flow cytometry analysis.The effect of Pygo2 over-expression on the level of cyclinD1 and β-catenin of C6 cells was detected by Western blot analysis,and the expression and subcellular location of cyclinD1 and β-catenin of C6 cells were further quantified by immunofluorescent staining.Results The recombinant plasmids were completely coincided with the designs by the restriction map and the sequence analysis,which up-regulated Pygo2 expression of C6 cells efficiently.After Pygo2 expression were up-regulated by transfected C6 cells with the recombinant plasmids,cells proliferation was promoted and colony forming was increased significantly,cell cycle progression from G1 to S transition was enhanced notably.Furthermore,the expression level of cyclinD1 was significantly increased without change of subcellular location,and the expression level and subcellular location of β-catenin were not changed obviously.Conclusion The recombinant vectors of Pygo2 over-expression were constructed successfully.Over-expression of Pygo2 promotes the growth of glioma cells by an increased expression of cyclinD1 to improve G1/S transition.重庆市自然科学基金资助项目(cstc2011jjA10110);重庆市教委科技基金资助项目(KJ100504);福建省自然科学基金资助项目(2009D002

湖泊底泥中微囊藻DNA的分子检测

通过改进裂解温度和延长裂解时间并增加苯酚/氯仿洗脱次数的DNA提取方法获得南京玄武湖底泥中的DNA,通过PCR法来扩增微囊藻的16SrRNA基因．结果表明在所有采样点中均得到微囊藻基因组DNA,并且纯度较高,OD_(260)/ OD_(280)均高于1．54,最高值达到1．89．PCR的扩增结果显示所有样点的DNA都得到212 bp大小的微囊藻16SrRNA基因片断,表明这种方法可以有效的从底泥中提取微囊藻的DNA,从而为研究底泥微囊藻生理生态及其越冬、上浮、形成水华的机理提供更有利的方法

影响中晚期肝细胞肝癌手术切除预后的多因素分析

探讨影响中晚期肝细胞肝癌手术切除预后的因素。方法对130例中晚期大肝癌随访1～7年,采用单因素、多因素分析统计不同预后因素对患者生存率的影响。结果手术后1,3 ,5年生存率分别81.7%,24.3%,18.4%。单因素分析提示影响预后的因素为肝癌大小、是否早期复发、肝硬化情况、输血量;多因素分析提示肝癌大小、肿瘤早期复发是影响肝癌术后的预后因素。结论：中晚期肝癌手术切除预后仍不理想,重视围手术期处理,预防术后早期复发有望提高手术疗效 [英文摘要]Objective To study the prognostic factors in patients who received hepatectomy for large hepatocellular carcinoma(HCC). Methods 130 patients operated for large HCC were followed up for 1～7 years. Twenty possible factors were analyzed by Kaplan-Meier Log rank estimate. A multivariative survival analysis of these individal variable was undertaken using the cumulative survival rate by the computers COX proportional hazard. Result The overall cumulative survival rate at 1,3,5 years was 81.7%,24.3%,18.4% re..

Zooplanktonic diversity in the western Pacific

西太平洋区是全球海洋生物种源中心,许多类群的最高物种多样性都出现于该区域,因此,在该区开展种类多样性的研究不仅重要和必要,而且具有在跨国尺度上进行综合管理和相互合作的迫切性。本文在西太平洋的浮游动物样品鉴定分类、编目、文献资料整理和分析的基础上,记录和编入西太平洋10个浮游动物类群2,658种(含亚种),隶属于206科675属,其中水螅水母类99科251属697种,栉水母类12科22属59种,浮游软体动物14科35属86种,介形类8科89属416种,桡足类51科156属908种,糠虾类4科58属202种,磷虾类2科8属56种,十足类8科22属105种,毛颚类5科8属48种,被囊类5科26属81种。The western Pacific region has been operating as a centre for the origin of marine biodiversity: the richest diversity of many marine taxa was found in these waters.Therefore,biodiversity research and con-servation efforts in this area are necessary in order to promote the integrative and international management of this resource.The present work is a compilation of numbers of all the families,genera and species of ma-jor taxa of zooplankton known in the western Pacific Ocean(106°–150°E,0°–44°N).In all,2,658 zooplank-tonic species(including subspecies) belonging to 206 families and 675 genera have been recorded from taxonomic identifications and literature,99 families,251 genera and 697 species belong to the Medusozoa,12 families,22 genera and 59 species to the Ctenophora,14 families,35 genera and 86 species to the pelagic Molluscs(Pteropoda and Heteropoda),8 families,89 genera and 416 species to the Ostracoda,51 families,156 genera and 908 species to the Copepoda,4 families,58 genera and 202 species to the Mysidcea,2 fami-lies,8 genera and 56 species to the Euphausiacea,8 families,22 genera and 105 species to the Decapoda,5 families,8 genera and 48 species to the Chaetognatha,5 families,26 genera and 81 species to the Tunicata.908专项“中国海洋生物种类名录和图谱”(908-ZC-II-02)和908专项海洋生物样品库;科技部基础工作专项“我国和邻近西北太平洋海洋生物物种编目和分布图集编制”(2006FY220700

Preparation and Electrochemical Lithium Intercalation Performance of Segmented Carbon Nanofibers

以泡沫镍为催化剂 ,在 6 0 0和 70 0℃下 ,以CVD法热解乙炔气体制备大量的纳米碳纤维 .随着制备温度增加 ,纳米碳纤维直径变小 ,竹节状含量减少 ,d0 0 2 值减小 ,微晶片层平面Lc 和La 值增大 ,碳材料的可逆容量则下降 .分别用透射电镜、X射线衍射和拉曼光谱观察和测定了纳米碳纤维的形貌、微结构 ,发现在不同条件下生长的纳米碳纤维有不同的形貌和结构 .对纳米碳纤维的电化学嵌锂性能的研究表明 ,纳米碳纤维的结构对其电化学嵌锂容量和充放电循环寿命起重要影响 ,制备温度越低 ,纳米碳纤维的石墨化程度越差 ,可逆嵌锂容量相应要高一些Segmented carbon nanofibers were prepared by pyrolysis of acetylene on foam Ni at 600 and 700℃ in a fixed bed flowed-reactor. The morphology, microstructure and lithium insertion properties of these carbon nanofibers were investigated by TEM, XRD, Raman and electrochemical methods. Through TEM observations, it was found that this kind of carbon nanofibers was composed of lens-like segments with nearly equal separation stacking along the nanofiber axis. When the reaction temperature was 600℃, segmented carbon nanofibers were the major production. However, when the reaction temperature increased to 700 ℃, the content of segmented carbon filaments decreased and their diameter became smaller. The crystallite size d 002 and L c were determined by the 002 carbon Bragg peak of XRD patterns using the Bragg and Scherrer formulas. The intensity ratios of the 1350 cm -1 line and the 1580 cm -1 line (R=I D/I G) was used to evaluate the L a value, which was inversely proportional to the effective crystallite size in the direction of the graphite plane (L a). With the reaction temperature increased, the d 002 value decreased, L a and L c values increased, which indicated the degree of crystallinity increased. Segmented carbon nanofibers were used as positive electrodes of C/Li cells. The first charge capacities of C/Li cells were 480 and 300 mAh/g for samples produced at 600 and 700℃, respectively. The samples at 600℃ showed capacities higher than the theoretical value of graphite, which was attributed to accommodation of more lithium at the edge of graphene layers and on the surface of graphene layers according to the mechanisms of lithium insertion in carbons prepared by low-temperature pyrolysis of hydrocarbons. As confirmed by the XRD and Raman spectra, the samples at 700℃ had larger L a and L c, which led to the capacity decreasing.国家自然科学基金 (6 0 2 710 0 9);; 浙江省自然科学基金 (5 0 110 9,2 0 0 0 5 3)资助项

一种定量检测人血清高敏C反应蛋白的化学发光免疫方法

旨在建立一种可定量检测人血清高敏CRP的化学发光检测方法(High-sensitivity C-reactive protein quantifiable chemiluminescent immunoassay,hs-CRP CLIA)。首先利用亲和层析和离子交换层析技术从肝硬化病人腹水中纯化出高纯度的天然CRP作为免疫原制备了22株CRP单克隆抗体(单抗),其中13株单抗在磷酸胆碱配体捕获ELISA中呈阳性,然后利用方正滴定法筛选出单抗10C5和10C11建立了hs-CRP CLIA。试剂盒评估结果显示:该方法对血清中干扰物质IgG、血红蛋白、甘油三酯等无非特异性反应;该方法检测灵敏度高,在0.04~20.38mg/L范围内定量检测人血清CRP标准品呈良好线性关系(R2>0.993);该方法准确性高、可重复性好,平均回收率为99%,批内差为4.2%~5.8%,批间差为9.0%~11.5%;该方法与进口商品化高敏CRP ELISA试剂盒平行比较检测90份血清标本,结果显示两者有良好的可比性(r=0.968)。综上,建立的hs-CRP CLIA是一种准确、可靠、可定量的高灵敏C反应蛋白检测方法,该方法的临床应用,有利于改善我国心脏病风险评估及肠炎性疾病预后判断